Alcohol oxidases are known to be produced by various microorganisms grown on methanol, though not on ethanol. These alcohol oxidases catalyze the reaction EQU RCH.sub.2 OH+O.sub.2 .revreaction.RCHO+H.sub.2 O.sub.2
where R is hydrogen or a lower alkyl, generally selected from the group H--, CH.sub.3 --, CH.sub.3 CH.sub.2 --, and CH.sub.3 (CH.sub.2).sub.2 --. Alcohol oxidases can be used to scavenge or remove oxygen from compatible solutions, as well as in the production of aldehydes and hydrogen peroxide. In combination with a suitable test probe, the alcohol oxidase enzyme can be employed to determine alcohol concentration, especially the concentrations of such alcohols as methanol and ethanol. Hence, such enzymes are useful in applications such as the measurement of alcohol levels in biological fluids, for example blood, and the like.
Some uses of these enzymes have been hindered by unavailability of the pure enzyme in commercially reasonable quantities for proposed uses. One of the problems encountered with previous types of alcohol oxidase is that, in common with many enzymes, they are difficult to isolate in relatively pure form, for example in a form essentially lacking catalase activity. A variety of techniques, such as fractional precipitation using materials such as ammonium sulfate, alcohol or polyethylene glycol, or column chromatography using ion exchange resins or gel molecular sieve media, have been utilized to prepare the purified enzyme. Isolation of relatively pure alcohol oxidase by the use of such techniques is difficult, and material and time consuming, accounting for much of the cost of the commercial enzyme. The resulting high costs of relatively pure alcohol oxidases lead to restricted usage of these enzymes, and discourage potential use of the enzyme in applications requiring large quantities of the enzyme as well as discouraging the search for new applications. Moreover, as is known in the art, stability of the known prior alcohol oxidase enzymes to temperature and pH, the specificity for particular substrates, and susceptibility to inhibition by these compounds and others, as well as the rate of the catalyzed reaction also affect the potential uses to which such enzymes are most efficiently and effectively placed.